Quantitative Determination of Alkaloids from Roots of Hydrastis canadensis L. and Dietary Supplements Using Ultra-Performance Liquid Chromatography with UV Detection
Quantitative Determination of Alkaloids from Roots of Hydrastis canadensis L. and Dietary Supplements Using Ultra-Performance Liquid Chromatography with UV Detection
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DOI：
10.5740/jaoacint.12-074
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发表时间：
2012-09-01
影响因子：
1.6
通讯作者：
Khan, Ikhlas A.
中科院分区：
文献类型：
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作者：
Avula, Bharathi;Wang, Yan-Hong;Khan, Ikhlas A.
采用超高效液相色谱(UPLC)结合紫外检测技术，对加拿大水蛇(Hydrastis canamsis L.)及声称含有黄花丹的膳食补充剂中的生物碱进行了定量测定。分析在Waters Acquity UPLC系统上进行，色谱柱为Acquity UPLC BEH Shield RP18，用甲酸铵和含有甲酸的乙腈进行梯度洗脱。色谱柱运行时间小于6min。对半胱氨酸和斑驳碱的检测波长为290 nm，对半胱氨酸、黄连碱、药根碱、巴马汀和黄连素的检测波长为344 nm。对100%甲醇、90%甲醇、90%甲醇+1%醋酸、90%乙腈+0.1%磷酸和100%乙腈5种不同的萃取剂进行了主要成分的回收试验。用90%甲醇+1%冰醋酸提取的样品回收率最高(97%)。对分析方法的线性、重复性、精密度和精密度进行了验证。日内和日间RSD均小于3.5%，回收率为98-103%。采用UPLC/MS、四极杆质谱仪和电喷雾电离源对7种生物碱进行了鉴定。该分析方法被成功地应用于加拿大黄连根部的7种生物碱、声称含有黄花的膳食补充剂以及可能的掺假物种的鉴定。
Ultra-performance liquid chromatography (UPLC) with UV detection was used for the quantification of alkaloids from roots of Hydrastis canadensis L. (goldenseal) and dietary supplements claiming to contain goldenseal. The analysis was performed on a Waters Acquity UPLC system with an Acquity UPLC BEH Shield RP18 column using gradient elution with ammonium formate and acetonitrile containing formic acid. The chromatographic run time was less than 6 min. The detection wavelength used for p-hydrastine and canadine was 290 nm; for hydrastinine, coptisine, jatrorrhizine, palmatine, and berberine, it was 344 nm. A total of five different extraction solvents, including 100% methanol, 90% methanol, 90% methanol + 1% acetic acid, 90% acetonitrile + 0.1% phosphoric acid, and 100% acetonitrile, were tested for recovery of the major compounds. The samples extracted with the 90% methanol + 1% acetic acid displayed the best recovery (>97%). The analytical method was validated for linearity, repeatability, LOD, and LOQ. The RSDs for intraday and interday experiments were less than 3.5%, and the recovery was 98-103%. UPLC/MS with a quadrupole mass analyzer and electrospray ionization source was used to confirm the identity of seven alkaloids. The analytical method was successfully applied to confirm the identification of seven alkaloids from the roots of H. canadensis, dietary supplements that claimed to contain goldenseal, and possible adulterant species.